Combinatorial methods for the synthesis of new drug substances require high throughput techniques for evaluation of structure-activity relationships. Microtiter plates with immobilized target receptors can rapidly reduce thousands of possibilities to hundreds of candidates. The next level of screening has proven to be more difficult and tedious. The goal of this project is creation of an on-line screening approach to expedite secondary screening. The new technology involves immobilized biopolymer HPLC columns (IBCs) containing immobilized receptors. Test compounds will be sequentially displaced using a characterized ligand and the liberated compounds directed on-line into a mass spectrometer. Thus, a large series of compounds can be rapidly screened for their relative affinities for the immobilized receptor with concurrent identification of molecular structures. The objective of this project is to develop prototypic IBC's. In phase I, immobilized nicotinic receptor-IBCs were developed and shown to bind test ligands. In phase II, the application of nicotinic receptor-IBCs to high-throughput screening will be demonstrated. PROPOSED COMMERCIAL APPLICATION: When applied to the drug discovery process, the described technology will facilitate the identification of new lead candidates. The immobilized receptor approach will improve efficiency in drug discovery because it will save time and money. The applicants have filed patents protecting the new technology and intend to offer it to companies with drug discovery as their mission. The sale of this technology to the pharmaceutical industry reflects the proposed commercial application on this project.